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Single Cell Protein Analysis

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Cover of 'Single Cell Protein Analysis'

Table of Contents

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    Book Overview
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    Chapter 1 Single-Cell Western Blotting
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    Chapter 2 A Microfluidic Device for Immunoassay-Based Protein Analysis of Single E. coli Bacteria
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    Chapter 3 Enzyme-Linked ImmunoSpot (ELISpot) for Single-Cell Analysis
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    Chapter 4 Single Cell Protein Analysis
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    Chapter 5 Single Cell Protein Analysis
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    Chapter 6 Microfluidic Flow Cytometry for Single-Cell Protein Analysis
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    Chapter 7 Microfluidic Image Cytometry for Single-Cell Phenotyping of Human Pluripotent Stem Cells
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    Chapter 8 Characterizing Phenotypes and Signaling Networks of Single Human Cells by Mass Cytometry.
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    Chapter 9 Multiplexed Peptide-MHC Tetramer Staining with Mass Cytometry.
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    Chapter 10 Imaging and Mapping of Tissue Constituents at the Single-Cell Level Using MALDI MSI and Quantitative Laser Scanning Cytometry
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    Chapter 11 SPLIFF: A Single-Cell Method to Map Protein-Protein Interactions in Time and Space.
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    Chapter 12 Microfluidic Proximity Ligation Assay for Profiling Signaling Networks with Single-Cell Resolution
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    Chapter 13 Dynamics and Interactions of Individual Proteins in the Membrane of Single, Living Cells
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    Chapter 14 Microfluidics-Enabled Enzyme Activity Measurement in Single Cells
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    Chapter 15 Microfluidic Chemical Cytometry for Enzyme Assays of Single Cells
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    Chapter 16 Quantitative Detection of Nucleocytoplasmic Transport of Native Proteins in Single Cells
Attention for Chapter 12: Microfluidic Proximity Ligation Assay for Profiling Signaling Networks with Single-Cell Resolution
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Chapter title
Microfluidic Proximity Ligation Assay for Profiling Signaling Networks with Single-Cell Resolution
Chapter number 12
Book title
Single Cell Protein Analysis
Published in
Methods in molecular biology, January 2015
DOI 10.1007/978-1-4939-2987-0_12
Pubmed ID
Book ISBNs
978-1-4939-2986-3, 978-1-4939-2987-0
Authors

Matthias Blazek, Günter Roth, Roland Zengerle, Matthias Meier

Abstract

The proximity ligation assay (PLA) is a technique that can be used to characterize proteins, protein-protein interactions, and protein modifications at the single-cell level. Image-based in situ detection of proteins using PLA is a quantitative method with a high degree of sensitivity and specificity. The miniaturization and parallelization of the PLA onto a microfluidic chip and concurrent use of an automated cell-culture system increase the throughput of this technology. Here, we describe the performance of PLA on a microfluidic chip. We provide protocols for on-chip cell culture, time-shifted cell stimulation and fixation, PLA implementation, and computational image analysis in order to achieve single-cell resolution. As a proof of concept, we studied the phosphorylation of Akt in response to stimulation with platelet-derived growth factor.

Mendeley readers

The data shown below were compiled from readership statistics for 10 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 10 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 3 30%
Professor > Associate Professor 2 20%
Student > Master 2 20%
Student > Postgraduate 1 10%
Student > Bachelor 1 10%
Other 0 0%
Unknown 1 10%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 4 40%
Immunology and Microbiology 2 20%
Agricultural and Biological Sciences 1 10%
Medicine and Dentistry 1 10%
Engineering 1 10%
Other 0 0%
Unknown 1 10%